Detection of human retrovirus infection

Abstract

A method for determining virus replication in human cells by human retrovirus using RNA amplification comprising detecting the hybridization of an RNA probe which specifically hybridizes with spliced RNA and not with genomic RNA. This method permits early detection of RNA replication resulting from primary infection without detecting non-replicating virus.

We claim: 
     
       1. A method for determining human immunodeficiency virus (HIV) replication resulting from primary infection of human cells using an RNA amplification method, comprising providing a sample of human peripheral blood lymphocytes, extracting the nucleic acid from the peripheral blood lymphocytes, adding amplification primers that hybridize with exon 1 and exon 2 of a spliced tat gene RNA segment, amplifying RNA comprising exon 1 and exon 2, adding a probe that will hybridize with the junction of exon 1 and exon 2 in a spliced RNA tat mRNA, and detecting the hybridization of said probe with said spliced RNA, wherein a positive result is obtained when hybridization occurs and indicates HIV replication. 
     
     
       2. The method of claim 1, wherein the probe is labeled with a detectable label selected from the group consisting of an enzyme, a sol particle and a radionuclide. 
     
     
       3. The method of claim 1, wherein the probe is detected using a labeled, complementary nucleic acid sequence that hybridizes specifically to the probe. 
     
     
       4. The method of claim 1, wherein the infection is detected within forty-eight hours of primary infection in-vitro. 
     
     
       5. A method for evaluating an antiviral agent or inhibiting antibody comprising introducing an antiviral agent or inhibiting antibody into serum containing human peripheral blood mononuclear cells and determining HIV replication by the method of claim 1. 
     
     
       6. The method of claim 5, wherein the antiviral agent or inhibiting antibody is evaluated by determining HIV replication within approximately the first forty-eight hours after infection in-vitro. 
     
     
       7. The method of claim 6, wherein the efficacy of the antiviral agent or inhibiting antibody is determined by obtaining the percent of inhibition of HIV infection. 
     
     
       8. The method of claim 1, wherein the peripheral blood lymphocytes are peripheral blood mononuclear cells.